embryo DNA protocol
byron_w14850 at yahoo.com
Thu Sep 5 13:51:30 EST 2002
We do DNA preps on single embryos for use in PCR reactions.
Adapted from Hatton & O'Hare 1999.
Pick the embryo off the grape plate with a toothpick and place it in a
clear 0.5 epp tube. Pop the embryo against the side (towards the
bottom preferably)-- visualize using dissecting scope.
Immediately add Extraction Buffer (10 mM Tris ph 8, 1 mM EDTA, 25 mM
NaCl, 1/100 dilution of 20 mg/ml Proteinase K.) Keep on ice until all
the embryos are done. Vortex all the tubes, then do a quick spin down.
Tubes in PCR machine for 37 degrees for 30 min, 95 degrees for 10
minutes. Spin down again. Use 1-1.5 ul as template in 1 ul reaction.
Hope this works.
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